Molecular identification of Vibrio harveyi-related isolates associated with diseased aquatic organisms

Gómez‐Gil, Bruno; Soto-Rodríguez, Sonia A.; García‐Gasca, Alejandra; Roque, Ana; Vázquez-Juárez, Ricardo; Thompson, Fabiano L.; Swings, Jean

doi:10.1099/mic.0.26797-0

Cited by 188 publications

(166 citation statements)

References 43 publications

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“…In recent years, sequencing and comparison of the 16S rRNA gene has become an important tool for identification of bacterial species (Greenwood et al, 2005). However, V. harveyi and V. campbellii share more than 97 % similarity in their 16S rRNA gene sequences and show a 69 % match in DNA-DNA hybridization (Gomez-Gil et al, 2004). Further, the wellstudied V. harveyi strain BB120 was recently proved to be V. campbellii by microarray-based comparative genomic hybridization (Lin et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Identification of Vibrio campbellii isolated from diseased farm-shrimps from south India and establishment of its pathogenic potential in an Artemia model

et al. 2011

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Shrimp diseases are frequently reported to be caused by closely related vibrios, and in many cases they are tentatively but inaccurately identified as Vibrio harveyi and related vibrios. In the present study, 28 biochemically identified V. harveyi-related strains isolated from diseased shrimps were randomly selected for further characterization by molecular tools. Twenty-six strains were identified as Vibrio campbellii and two as V. harveyi by sequence analysis of 16S rRNA and uridylate kinase genes. Haemolysin-gene-based species-specific multiplex PCR also confirmed these results. Experimental challenge studies using Artemia as a model showed that eight isolates were highly pathogenic, three were moderately pathogenic and the remaining 17 were non-pathogenic. Ribotyping with BglI clearly distinguished V. campbellii from V. harveyi, but it failed to separate pathogenic and non-pathogenic clusters. Artemia nauplii challenged with a fluorescently labelled highly pathogenic strain (IPEY54) showed patches in the digestive tract. However, no patches were observed for a non-pathogenic strain (IPEY41). Direct bacterial counts also supported colonization potential for the highly pathogenic strain. To our knowledge, this is the first report on the isolation and accurate identification of large numbers of V. campbellii associated with shrimp disease in aquacultural farms. V. campbellii has long been considered to be non-pathogenic and classified with V. harveyi-related bacteria. However, we show that this species may be an emerging aquaculture pathogen. This study will help to formulate suitable strategies to combat this newly identified pathogen. INTRODUCTIONHalophilic vibrios such as Vibrio harveyi are ubiquitous in the marine environment and are implicated as the causes of several diseases in wild and cultured aquatic organisms. Due to the plasticity of Vibrio genomes, with frequent horizontal gene transfer events, species boundaries are very narrow in the marine environment (Fraser et al., 2007). Hence, the identification of V. harveyi and related species isolated from the marine environment is sometimes difficult. Luminous vibrios including V. harveyi have been implicated principally with disease outbreaks in shrimp larval culture facilities and grow-out ponds worldwide. Due to the high level of phylogenetic similarity among marine vibrios, bacteria associated with disease outbreaks have often been misidentified. For example, although strain LMG 19703 T (or AK1 T ) showed 99.4 % sequence similarity in 16S rRNA with Vibrio mediterranei (ATCC 43341 T ), Kushmaro et al. (2001) initially classified it as a new species, Vibrio shiloi, due to large differences in phenotypic properties. Later, Thompson et al. (2001), on the basis of genotypic features such as fluorescent amplified fragment length polymorphism and DNA-DNA hybridization, as Abbreviations: ASW, artificial seawater; DOC, days of culture; DTAF, 5-(4,6-dichlorotriazin-2-yl)aminofluorescein; NJ, neighbour joining. The GenBank/EMBL/DDBJ accession numbers for the se...

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Section: Introductionmentioning

confidence: 99%

Identification of Vibrio campbellii isolated from diseased farm-shrimps from south India and establishment of its pathogenic potential in an Artemia model

et al. 2011

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“…Its fisheries have been performed mainly in the East Sea over several decades [7]. Vibrio (V.) harveyi belonging to the genus Vibrio has been reported to infect a wide range of marine organisms worldwide including fish and invertebrates [1,3,6]. The disease can cause mass mortalities in aquaculture species with significant economic impact.…”

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confidence: 99%

Isolation and identification of Vibrio harveyi from chub mackerel (Scomber japonicus)

Lee¹,

Jun²,

Giri³

et al. 2016

Korean Journal of Veterinary Research

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: For several days, there was a series of mortalities of chub mackerel (Scomber japonicus) that were reared for public exhibition in a private aquarium in Seoul, Korea. As part of the diagnosis of the dead fish, a bacterial isolate from the kidney was cultured, identified, and confirmed to be Vibrio (V.) harveyi using Vitek System 2 and 16S rRNA gene sequencing. Phylogenetic analysis was also performed by the neighbor-joining method. As a result, the V. harveyi isolated from chub mackerels of a private aquarium in Korea, called as SNUVh-LW1, was clustered in the same group with V. harveyi ATCC33843.

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“…In the family Vibrionaceae, sequence similarities for the 16S rRNA gene are ≥ 97.6% among members of the so-called Vibrio core group, which is formed by 6 species, V. harveyi, V. campbellii, V. ro ti ferianus, V. parahaemolyticus, V. algino ly ticus and V. natriegens (Dorsch et al 1992). Recent studies have shown that the 16S rRNA gene is not adequate for resolution of the 6 species of the core group of the genus Vibrio, because the overlap between the intraspecific and interspecific distances is very large (Gomez-Gil et al 2004, Lin et al 2010, Pascual et al 2010. The phylogenetic analysis of the 16S rRNA gene of strain AP37 also confirms this conclusion.…”

Section: Discussionmentioning

confidence: 99%

Identification and characterization of Vibrio harveyi associated with diseased abalone Haliotis diversicolor

Jiang¹,

Shi²,

Ke³

et al. 2013

Dis. Aquat. Org.

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Mass mortality of farmed small abalone Haliotis diversicolor occurred in Fujian, China, from 2009 to 2011. Among isolates obtained from moribund abalones, the dominant species AP37 exhibited the strongest virulence. After immersion challenge with 10 6 CFU ml −1 of AP37, abalone mortalities of 0, 53 and 67% were induced at water temperatures of 20°C, 24°C, and 28°C, respectively. Following intramuscular injection, AP37 showed a low LD 50 (median lethal concentration) value of 2.9 × 10 2 CFU g −1 (colony forming units per gram abalone wet body weight). The LT 50 (median lethal time) values were 5.2 h for 1 × 10 6 CFU abalone . For further analysis of virulence, AP37 was screened for the production of extracellular factors. The results showed that various factors including presence of flagella and production of extracellular enzymes, such as lipase, phospholipase and haemolysin, could be responsible for pathogenesis. Based on its 16S rRNA gene sequence, strain AP37 showed > 98.8% similarity to Vibrio harveyi, V. campbellii, V. parahaemolyticus, V. algi nolyticus, V. na trie gens and V. rotiferianus, so it could not be identified by this method. However, multi-locus sequence analysis (MLSA) of concatenated sequences, including the rpoD, rctB, gyrB, toxR and pyrH genes, identified strain AP37 as V. harveyi. Phenotypic characters of AP37 were identified by API 20E. In antibiotic susceptibility tests, strain AP37 exhibited susceptibility to 7 antibiotics and resistance to 13. This is the first report of a V. harveyi-related species being linked with the mass mortality of adult abalone H. diversicolor in southern China.

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Molecular identification of Vibrio harveyi-related isolates associated with diseased aquatic organisms

Cited by 188 publications

References 43 publications

Identification of Vibrio campbellii isolated from diseased farm-shrimps from south India and establishment of its pathogenic potential in an Artemia model

Identification of Vibrio campbellii isolated from diseased farm-shrimps from south India and establishment of its pathogenic potential in an Artemia model

Isolation and identification of Vibrio harveyi from chub mackerel (Scomber japonicus)

Identification and characterization of Vibrio harveyi associated with diseased abalone Haliotis diversicolor

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